CN110004023B - Centrifugal microfluidic chip and nucleic acid analysis system - Google Patents
Centrifugal microfluidic chip and nucleic acid analysis system Download PDFInfo
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- CN110004023B CN110004023B CN201910130950.9A CN201910130950A CN110004023B CN 110004023 B CN110004023 B CN 110004023B CN 201910130950 A CN201910130950 A CN 201910130950A CN 110004023 B CN110004023 B CN 110004023B
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Abstract
The application relates to a centrifugal microfluidic chip and a nucleic acid analysis system, wherein the centrifugal microfluidic chip comprises a chip substrate, a magnetic absorption part and a magnetic part; the chip substrate is also provided with a stirring chamber and a liquid injection hole communicated with the stirring chamber; the magnetic absorption part is accommodated in the stirring chamber; the chip substrate has a rotation center, and the magnetic piece is arranged outside the chip substrate and positioned at a position of the stirring cavity close to the rotation center. On the one hand, when the magnetic adsorption piece moves to a position far away from the magnetic piece, the magnetic adsorption piece moves to a position of the stirring cavity far away from the rotation center under the action of centrifugal force; on the other hand, when the magnetic adsorption piece moves to a position close to the magnetic piece, the magnetic adsorption piece moves to a position of the stirring cavity close to the rotation center under the action of magnetic adsorption; therefore, the ideal mixing effect is realized, and the cracking effect is stable and effective on the premise of simplifying the structure and needing no complex instruments and equipment.
Description
Technical Field
The application relates to the field of centrifugal microfluidics, in particular to a centrifugal microfluidic chip and a nucleic acid analysis system.
Background
Microfluidic refers to manipulation of liquids on a sub-millimeter scale, where sub-millimeter scale is typically a few micrometers to hundreds of micrometers. Microfluidic technology integrates basic operating units involved in the biological and chemical fields, even the functions of the whole laboratory, including sampling, dilution, reaction, separation, detection, etc., on a small Chip, so called Lab-on-a-Chip. The chip is generally composed of various liquid reservoirs and a micro-channel network which are connected with each other, so that the sample processing time can be shortened to a great extent, and the maximum utilization efficiency of reagent consumable materials can be realized by precisely controlling the liquid flow.
Microfluidic systems refer to devices that manipulate liquids on a sub-millimeter scale. Centrifugal microfluidics belongs to one branch of microfluidics, and particularly relates to a centrifugal microfluidic chip which is rotated to drive the flow of liquid, so that the liquid is controlled on a submillimeter scale by using centrifugal force. Centrifugal microfluidics integrates basic operating units involved in the biological and chemical fields on a small disc-shaped (disk-shaped) chip. Besides the advantages peculiar to the microfluidic, the whole device is more compact and compact since the centrifugal microfluidic requires only one motor to provide the force required for liquid manipulation. The ubiquitous centrifugal field on the disc chip can not only enable liquid to be driven more effectively and ensure that no residual liquid exists in the pipeline, but also effectively realize sample separation based on density difference and enable parallel processing to be simpler. Microfluidic provides a very broad prospect for application in a plurality of fields such as biomedical research, drug synthesis screening, environmental monitoring and protection, health quarantine, judicial identification, detection of biological reagents and the like. In particular, microfluidic is well suited to the demands of Point-of-care testing (POCT) miniaturized instruments, and thus is widely used in POCTs. In industrialization, microfluidics is generally divided into the following large categories: pressure (pneumatic or hydraulic) driven microfluidic, centrifugal microfluidic, droplet microfluidic, digital microfluidic, paper microfluidic, etc.
In the microfluidic, the volume of liquid is in the microliter or even nanoliter level, and the sizes of the chamber and the pipeline of the microfluidic chip are limited, so that the mixing of reagents in the microfluidic chip is difficult to realize. In centrifugal microfluidic, there are two most common ways of mixing liquid reagents. One way is to realize the mixing effect by changing the rotational speed of the microfluidic chip and using the euler force generated in the acceleration and deceleration process of the microfluidic chip. Second, the mixing effect is achieved by the liquid flowing through a serpentine flow conduit that is continuously bent. The mixing of the first mode often puts a larger requirement on a centrifugal motor, and meanwhile, as the rotating speed in the centrifugal microfluidic is the power for controlling the liquid flow on the whole chip, the change of the rotating speed can have influence on the flow of liquid reagents at other parts on the microfluidic chip, and the design difficulty of the chip is increased. The second approach increases the difficulty of processing the entire microfluidic chip, while the mixing effect of the serpentine flow channel, which is constantly bent, is often less than ideal.
In addition, a very large block of application scenario of the whole microfluidic technology is microfluidic molecular diagnostics. The core purpose is to enrich, crack, extract and purify nucleic acid of the whole sample of molecular diagnosis, PCR amplification is integrated on a microfluidic chip. Here, the lysis of the sample has been a major technical difficulty in the diagnosis of the entire microfluidic molecule. Cleavage of samples in molecular diagnostics is generally divided into physical and chemical methods. For chemical methods, enzymes are typically used to hydrolyze cell membranes or cell walls, but the components of the reagents required for lysis for different pathogens are different, making the method less versatile. On the other hand, chemical methods require a long time for cleavage, and many times the cleavage effect is not ideal. Physical lysis typically uses mechanical means to disrupt the sample cell membrane or cell wall, thereby releasing DNA. The physical method generally includes an ultrasonic method, a laser irradiation method, and the like. In centrifugal microfluidics, these physical methods tend to greatly increase the complexity of the instrument. In addition, in the operation process, the micro-fluidic chip is in a centrifugal state, so that the ultrasonic probe is difficult to directly contact with the micro-fluidic chip, and the cracking effect is difficult to reach expectations. On the other hand, the laser irradiation method is used for cracking, and the light spot of the laser is difficult to align with the cracking cavity of the microfluidic chip.
Disclosure of Invention
Based on this, it is necessary to provide a centrifugal microfluidic chip and a nucleic acid analysis system.
A centrifugal microfluidic chip, comprising: the chip comprises a chip substrate, a magnetic absorption part and a magnetic part; the chip substrate is also provided with a stirring chamber and a liquid injection hole communicated with the stirring chamber; the magnetic absorption part is accommodated in the stirring cavity; the chip substrate is provided with a rotation center, the magnetic piece is arranged outside the chip substrate and positioned at the position of the stirring cavity close to the rotation center, and the magnetic piece is used for enabling the magnetic absorption piece to move to the position of the stirring cavity close to the rotation center under the action of magnetic absorption when the distance between the magnetic piece and the magnetic absorption piece is smaller than a preset value.
According to the centrifugal microfluidic chip, the smart design that the magnetic absorption part is movably arranged in the stirring cavity is adopted, and on one hand, when the magnetic absorption part moves to a position far away from the magnetic part, the magnetic absorption part moves to a position far away from the rotation center of the stirring cavity under the action of centrifugal force; on the other hand, when the magnetic adsorption piece moves to a position close to the magnetic piece, the magnetic adsorption piece moves to a position of the stirring cavity close to the rotation center under the action of magnetic adsorption; therefore, the ideal mixing effect is realized, and the cracking effect is stable and effective on the premise of simplifying the structure and needing no complex instruments and equipment.
In one embodiment, the magnetic member is a magnetic body and the magnetic attachment is a ferromagnetic body that matches the magnetic body.
In one embodiment, the ferromagnetic body is provided with a protective outer layer.
In one embodiment, the magnetic member is a ferromagnetic body and the magnetic attachment member is a magnetic body that mates with the ferromagnetic body.
In one embodiment, the centrifugal microfluidic chip further comprises an abrasive body disposed in the agitation chamber.
In one embodiment, the centrifugal microfluidic chip further comprises a support member disposed on the chip base body and not rotating with the chip base body, and the magnetic member is fixedly disposed on the support member.
In one embodiment, the magnetic element is arranged separately from the chip base body.
In one embodiment, the number of the magnetic absorbing parts and the stirring chambers is plural, and each stirring chamber accommodates one magnetic absorbing part.
In one embodiment, the shape of each stirring chamber is the same or different.
A nucleic acid analysis system comprising the centrifugal microfluidic chip of any one of claims.
According to the nucleic acid analysis system, the ingenious design that the magnetic adsorption piece is movably arranged in the stirring cavity is adopted, and on one hand, when the magnetic adsorption piece moves to a position far away from the magnetic piece, the magnetic adsorption piece moves to a position far away from the rotation center of the stirring cavity under the action of centrifugal force; on the other hand, when the magnetic adsorption piece moves to a position close to the magnetic piece, the magnetic adsorption piece moves to a position of the stirring cavity close to the rotation center under the action of magnetic adsorption; therefore, the ideal mixing effect is realized, and the cracking effect is stable and effective on the premise of simplifying the structure and needing no complex instruments and equipment.
Drawings
Fig. 1 is a schematic external view of an embodiment of a centrifugal microfluidic chip according to the present application.
FIG. 2 is another schematic view of the embodiment of FIG. 1.
FIG. 3 is another schematic view of the embodiment of FIG. 1.
Fig. 4 is a sectional view in the A-A direction of the embodiment shown in fig. 3.
Fig. 5 is a schematic external view of another embodiment of a centrifugal microfluidic chip according to the present application.
Detailed Description
In order to make the above objects, features and advantages of the present application more comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. This application is, however, susceptible of embodiment in many other forms than those described herein and similar modifications can be made by those skilled in the art without departing from the spirit of the application, and therefore the application is not to be limited to the specific embodiments disclosed below.
It will be understood that when an element is referred to as being "fixed" or "disposed" on another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and the like are used herein for illustrative purposes only and are not meant to be the only embodiment.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
In the microfluidic, the volume of liquid is in the microliter or even nanoliter level, and the sizes of the chamber and the pipeline of the microfluidic chip are limited, so that the mixing of reagents in the microfluidic chip is difficult to realize. In one embodiment of the present application, a centrifugal microfluidic chip, comprising: the chip comprises a chip substrate, a magnetic absorption part and a magnetic part; the chip substrate is also provided with a stirring chamber and a liquid injection hole communicated with the stirring chamber; the magnetic absorption part is accommodated in the stirring cavity; the chip substrate is provided with a rotation center, the magnetic piece is arranged outside the chip substrate and positioned at the position of the stirring cavity close to the rotation center, and the magnetic piece is used for enabling the magnetic absorption piece to move to the position of the stirring cavity close to the rotation center under the action of magnetic absorption when the distance between the magnetic piece and the magnetic absorption piece is smaller than a preset value. According to the centrifugal microfluidic chip, the smart design that the magnetic absorption part is movably arranged in the stirring cavity is adopted, and on one hand, when the magnetic absorption part moves to a position far away from the magnetic part, the magnetic absorption part moves to a position far away from the rotation center of the stirring cavity under the action of centrifugal force; on the other hand, when the magnetic adsorption piece moves to a position close to the magnetic piece, the magnetic adsorption piece moves to a position of the stirring cavity close to the rotation center under the action of magnetic adsorption; therefore, the ideal mixing effect is realized, and the cracking effect is stable and effective on the premise of simplifying the structure and needing no complex instruments and equipment.
In one embodiment, the preset value, i.e. the preset distance, is set according to a balance relationship between the magnetic force of the magnetic field and the centrifugal force of the centrifugal field. In one embodiment, the magnetic member is configured to move the magnetic attraction member to a position of the stirring chamber near the rotation center under the magnetic attraction effect when the distance between the magnetic member and the magnetic attraction member is smaller than a preset value, that is, the magnetic member is configured to cooperate with the magnetic attraction member to move the magnetic attraction member to a position of the stirring chamber near the rotation center under the magnetic attraction effect. In one embodiment, the magnetic piece and the magnetic adsorption piece are selected according to the balance relation between the magnetic force of the magnetic field and the centrifugal force of the centrifugal field, when the distance between the stirring chamber and the magnetic piece is greater than the preset distance, the magnetic adsorption piece moves to the position of the stirring chamber away from the rotation center under the action of the centrifugal force of the centrifugal field, and when the distance between the stirring chamber and the magnetic piece is smaller than the preset distance, the magnetic adsorption piece moves to the position of the stirring chamber close to the rotation center, namely the position close to the magnetic piece, under the action of the magnetic force of the magnetic field, so that the stirring and mixing effects of liquid in the stirring chamber are realized.
In one embodiment, the magnetic attraction member is movably accommodated in the stirring chamber; in one embodiment, the magnetic attraction member is slidingly and/or rollingly received in the agitation chamber. In one embodiment, the magnetic attachment is a sphere that is received in the stir chamber and is capable of sliding and rolling movement in the stir chamber. The design is favorable for realizing the moving effect of the magnetic adsorption piece, thereby realizing more ideal mixing effect. Further, in one embodiment, the stirring chamber is further provided with a cover part, and the cover part is used for closing the liquid injection hole, and the liquid injection hole is used for injecting liquid. Further, in one embodiment, the stirring chamber is further provided with a liquid outlet hole and a sealing part thereof, and the sealing part is used for sealing the liquid outlet hole; in one embodiment, the liquid outlet hole and the liquid injection hole are respectively positioned on two sides of the chip substrate; after the stirring is finished, taking out the chip matrix, turning over the chip matrix until the liquid outlet is upward, opening the sealing part, connecting the liquid outlet pipe, turning over the chip matrix until the liquid injection hole is upward, and opening the sealing part to realize the outflow of the sample; and such a design is advantageous for cleaning the centrifugal microfluidic chip.
In order to avoid that the synchronous rotation of the magnetic element and the chip base body results in influencing the mixing effect, in one embodiment the magnetic element is arranged separately from the chip base body. In one embodiment, the magnetic element is arranged in a rotationally non-synchronized manner with the chip base body. In one embodiment, the magnetic member is in sliding contact with the chip substrate. In one embodiment, the centrifugal microfluidic chip further comprises a support member disposed on the chip base body and not rotating with the chip base body, and the magnetic member is fixedly disposed on the support member. In various embodiments, the magnetic member is configured to remain in place as the chip substrate rotates. The design is favorable for the magnetic adsorption piece in the stirring cavity to move to the position of the stirring cavity close to the rotation center under the magnetic adsorption effect once when the chip substrate rotates for one circle, namely, a mixing effect is generated. In one embodiment, the magnetic member is cylindrical. In one embodiment, the magnetic member is a rectangular body. It will be appreciated that in other embodiments, the shape of the magnetic member is not limited and it is only necessary to be able to magnetically attract the magnetic member to overcome the centrifugal effect and move from a position where the stirring chamber is away from the rotation center to a position where the stirring chamber is close to the rotation center. In one embodiment, the magnetic attraction member comprises a sphere. Alternatively, in one embodiment, the magnetic attraction member comprises a cylindrical shape.
In one embodiment, the number of the magnetic absorption parts and the stirring chambers are multiple, and each stirring chamber accommodates one magnetic absorption part; that is, the liquid injection holes are also multiple, and each liquid injection hole is correspondingly communicated with a stirring cavity; it is also understood that the plurality of liquid injection holes, the plurality of magnetic absorption parts and the plurality of stirring chambers are arranged in a one-to-one correspondence. In one embodiment, the number of the magnetic adsorbing elements and the stirring chambers is 2, 3, 4, 5, 6, 7, 8 or 9. In one embodiment, the shape of each stirring chamber is the same or different. The design of a plurality of stirring chambers is beneficial to simultaneously processing a plurality of samples; the stirring chambers have the same shape, which is beneficial to processing the same or different samples under the same condition or processing a plurality of samples under the same condition; the different shapes of the stirring chambers are beneficial to processing the same or different samples under different conditions, especially processing the same sample under different conditions, so that various different research modes and large-scale processing modes can be realized. In one embodiment, a plurality of the stirring chambers are symmetrically arranged with respect to the rotation center. In one embodiment, each stirring chamber is identical in shape, and a plurality of stirring chambers are symmetrically arranged relative to the rotation center. In one embodiment, the stirring chambers are identical in shape, and each stirring chamber is symmetrically arranged with respect to the rotation center.
In order to facilitate the cooperation to realize the rotation of the centrifugal microfluidic chip, in one embodiment, the centrifugal microfluidic chip is provided with a connecting portion or a mounting portion, and the connecting portion or the mounting portion is used for positioning and mounting the centrifugal microfluidic chip. In one embodiment, the number of the connection portions or the mounting portions is plural. In one embodiment, the mounting portion includes a mounting blind square hole or a mounting groove, or the like. In order to facilitate the rotation of the centrifugal microfluidic chip in cooperation, in one embodiment, the centrifugal microfluidic chip is provided with a rotation axis. In one embodiment, the centrifugal microfluidic chip is provided with a rotation shaft at a central position thereof. In one embodiment, the centrifugal microfluidic chip is of a central symmetrical structure, and the centrifugal microfluidic chip is provided with a rotation shaft or a rotation base at the symmetrical center thereof. In one embodiment, the stir chamber comprises a bar shape. In one embodiment, the rotation center is located in the direction of extension of the stirring chamber, i.e. the direction of extension of the stirring chamber in the form of a bar passes through the rotation center. By means of the design, when the centrifugal microfluidic chip rotates, the magnetic absorption piece can conveniently move from one end of the strip-shaped stirring cavity to the other end of the strip-shaped stirring cavity. In one embodiment, the rotation center is located on one side of the direction of extension of the stirring chamber, i.e. the direction of extension of the stirring chamber in the form of a bar does not pass through the rotation center. In one embodiment, the stir chamber comprises an arc and/or an ellipsoid. The shape and the position of the stirring cavity can be flexibly designed according to the requirements, and the magnetic adsorption piece can conveniently move from the position of the stirring cavity, which is close to the rotation center, to the position of the stirring cavity, which is far away from the rotation center, when the stirring cavity is close to the magnetic piece, and can move from the position of the stirring cavity, which is far away from the rotation center, to the position of the stirring cavity, which is close to the rotation center, only when the stirring cavity is close to the magnetic piece.
In one embodiment, the magnetic member is a magnetic body and the magnetic attachment is a ferromagnetic body that matches the magnetic body; the magnetic piece is used for generating magnetic adsorption force on the magnetic adsorption piece in the stirring cavity when the centrifugal microfluidic chip is in a rotating state and the stirring cavity is in a state of approaching the magnetic piece from far to near, so that the magnetic adsorption piece moves to a position of the stirring cavity, which is close to the rotating center, under the magnetic adsorption effect. In one embodiment, the magnetic member is a magnetic body and the magnetic attachment is a ferromagnetic body that matches the magnetic body. In one embodiment, the magnetic member is a cylindrical magnetic body and the magnetic attachment is a spherical ferromagnetic body or a cylindrical ferromagnetic body that matches the magnetic body. In one embodiment, the magnetic attraction member is provided with a protective outer layer. This is advantageous for protecting the magnetic attraction member to extend its service life. In one embodiment, the ferromagnetic body is provided with a protective outer layer. In one embodiment, the protective outer layer is a stainless steel layer or a ceramic layer for rust protection. In one embodiment, the ferromagnetic body includes Fe, co, ni elements and alloys thereof, rare earth elements and alloys thereof, and some Mn compounds. The magnetic body is an object generating a magnetic field; the strong magnet can react to the magnetic field to realize matching with the magnetic body, so that the magnetic absorption part moves to a position of the stirring cavity close to the rotation center under the action of magnetic absorption; that is, when the centrifugal microfluidic chip rotates, the magnetic adsorption piece moves to a position where the stirring chamber is far away from the rotation center due to the effect of centrifugal force, and when the stirring chamber is close to the position of the magnetic piece, the magnetic adsorption piece movably arranged in the stirring chamber moves towards the magnetic piece under the effect of magnetic adsorption, so that the magnetic adsorption piece moves to a position where the stirring chamber is close to the rotation center, and further stirring, namely mixing, inside the stirring chamber is realized. Further, in one of the embodiments, the magnetic member is a cylindrical magnetic body and the magnetic absorbing member is a spherical ferromagnetic body matching the magnetic body. In one embodiment, the spherical ferromagnetic body comprises a spherical ferromagnetic body and/or an ellipsoidal ferromagnetic body. In one embodiment, the magnetic member is a ferromagnetic body and the magnetic attachment member is a magnetic body that mates with the ferromagnetic body. In one embodiment, the magnetic member is a cylindrical ferromagnetic body and the magnetic attachment member is a spherical magnetic body that mates with the cylindrical ferromagnetic body. In one embodiment, a ferromagnetic material block, such as an iron block or a magnet block, on or in a stirring cavity of the centrifugal microfluidic chip is moved back and forth by utilizing a balance relationship between magnetic force of a magnetic field and centrifugal force of a centrifugal field, so that stirring and mixing effects of liquid in the stirring cavity are realized.
To enhance the lysing effect, in one embodiment, the centrifugal microfluidic chip further comprises a grinder disposed in the agitation chamber. Further, in one of the embodiments, the abrasive body comprises a spherical or ellipsoidal shape. In one embodiment, the abrasive body comprises ceramic beads, glass beads, quartz sand, diamagnetic spheres, diamagnetic ellipsoids, ferromagnetic stainless steel spheres, or ferromagnetic stainless steel ellipsoids. In one embodiment, the number of abrasive bodies is a plurality. In one embodiment, the volume ratio of the grinding body to the magnetic absorption piece is used for further improving the cracking effect, in one embodiment, the magnetic absorption piece is a spherical strong magnet or a spherical ferromagnetic body, the spherical strong magnet or the spherical ferromagnetic body is provided with a protruding part, and in one embodiment, the protruding part comprises a part-spherical protruding part, a conical protruding part and/or a conical protruding part; in one embodiment, the part-spherical protrusion is a hemispherical protrusion. Further, in one embodiment, the design is beneficial to having a certain cracking effect under the premise of simplifying the structure and not needing complex instruments and equipment, and especially has better cracking effect when being matched with the design of the grinding body. The current application scene of the whole microfluidic technology is microfluidic molecular diagnosis, and the core purpose of the method is to integrate the whole sample enrichment, cracking, nucleic acid extraction and purification of the molecular diagnosis and PCR amplification on a microfluidic chip structure. Among them, the sample is always cracked as a big difficulty in the whole microfluidic molecular diagnosis technology. In one embodiment, on the basis of the back and forth vibration, stirring and mixing of the ferromagnetic substance blocks, glass beads or quartz sand are preset in the stirring chamber in one embodiment, and the repeated back and forth movement of the ferromagnetic substance blocks in the stirring chamber can play a role in grinding, so that cell membranes or cell walls are broken to release DNA or RNA, and further the cracking of a sample by a physical method is realized.
In one embodiment, the stirring chamber comprises an arc and/or ellipsoid shape; the magnetic adsorption piece is provided with a protective outer layer; the centrifugal microfluidic chip further comprises a grinding body arranged in the stirring cavity; the magnetic piece is arranged separately from the chip substrate.
By the design of each embodiment, the ferromagnetic material blocks in the stirring cavity on the centrifugal microfluidic chip are enabled to move back and forth by utilizing the balance relation between the magnetic force of the magnetic field and the centrifugal force of the centrifugal field, so that the stirring and mixing effects of liquid in the stirring cavity are realized. In one embodiment, as shown in fig. 1 to 4, at least one agitation chamber 130 is provided on the centrifugal microfluidic chip 110. A liquid injection hole 150 is formed in the stirring chamber 130 near the center of the centrifugal center, a liquid sample is injected through the liquid injection hole 150, and a ferromagnetic substance block 140 serving as the magnetic absorption member is placed in the stirring chamber 130. A permanent magnet 160 is fixed below the centrifugal microfluidic chip as the magnetic member, and the center of the permanent magnet 160 is closer to the center of the circle than the center of the stirring chamber 130. Thus, when the centrifugal microfluidic chip rotates to the stirring chamber 130 to be close to the permanent magnet 160, for example, above the permanent magnet 160, the ferromagnetic substance 140 is attracted to the end of the stirring chamber 130 close to the center of the circle under the action of the magnetic field of the permanent magnet 160. When the centrifugal microfluidic chip rotates to a distance between the stirring chamber 130 and the fixed permanent magnet 160, the magnetic force is smaller than the centrifugal force, and the ferromagnetic material block 140 is thrown to the end of the stirring chamber 130 away from the center of the centrifugal force. Thus, when the centrifugal microfluidic chip rotates at a low speed, the ferromagnetic substance block 140 moves back and forth in the stirring chamber 130, so as to realize a stirring and mixing effect on the liquid in the stirring chamber 130, i.e. the sample. Further, if a grinding body such as glass beads or quartz sand is preset in the stirring chamber 130, mechanical lysis of cells, bacteria or viruses, etc. in the sample in the stirring chamber 130 can be better achieved. When the rotational speed of the centrifugal microfluidic chip increases, the speed at which the ferromagnetic substance block 140 moves back and forth in the stirring chamber 130 increases, and the stirring effect and the lysis effect are better. However, when the rotational speed of the centrifugal microfluidic chip increases to a certain value, even when the stirring chamber 130 is directly above the permanent magnet 160, the magnetic force is insufficient to attract the ferromagnetic substance block 140 to the end of the stirring chamber 130 chamber near the center of the circle against the centrifugal force. The ferromagnetic pieces 140 cannot move back and forth in the stirring chamber 130, and stirring and cracking are disabled; this rotational speed is therefore limited. Ferromagnetic pieces 140 include, but are not limited to, magnet pieces or iron pieces, and the like. The magnetic absorption part in the stirring chamber 130 may be a permanent magnet, and a common iron block is fixed outside the stirring chamber 130, i.e. outside the chip substrate. Further, in one embodiment, a plurality of stirring chambers 130 are designed at the same radius of the centrifugal microfluidic chip, and a ferromagnetic block 140 is placed in each stirring chamber, so that when the centrifugal microfluidic chip rotates, the stirring/cracking effect of the plurality of stirring chambers can be realized. As shown in fig. 4, 4 symmetrical stirring chambers are designed at the same radius of the centrifugal microfluidic chip, which can also be called as cracking stirring chambers, and when the centrifugal microfluidic chip rotates, stirring/cracking operation can be realized on the 4 stirring chambers at the same time; correspondingly, as shown in fig. 1, the centrifugal microfluidic chip 110 is provided with 4 liquid injection holes 150 respectively communicated with 4 stirring chambers, and each liquid injection hole is correspondingly communicated with one stirring chamber. As shown in fig. 2 and 3, the permanent magnet 160 is disposed in sliding contact with the chip base 110, i.e., the magnetic member is in sliding contact with the chip base. Alternatively, as shown in fig. 5, the permanent magnet 160 is provided separately from the chip base 110, that is, the magnetic member is provided separately from the chip base. By means of the design, the ferromagnetic material blocks in the stirring cavity on the centrifugal microfluidic chip are enabled to move back and forth by means of the balance relation between magnetic force of the magnetic field and centrifugal force of the centrifugal field, so that the stirring and mixing effects of liquid in the stirring cavity are achieved, and mechanical cracking of cells, bacteria, viruses and the like in samples in the stirring cavity can be achieved by further matching with glass beads or quartz sand preset in the stirring cavity; the design of the stirring and cracking modes is very simple to realize, only a permanent magnet is needed to be fixed below the centrifugal microfluidic chip, and a ferromagnetic substance block is preset in the stirring/cracking cavity, the cracking/stirring can be realized by only controlling the rotation speed of the centrifugal microfluidic chip, and the cracking/stirring effect can be optimized by adjusting the rotation speed; the cracking/stirring effect can be stopped by further increasing the rotating speed of the centrifugal micro-fluidic chip on the basis; and the parallel cracking/stirring effect of a plurality of stirring chambers can be realized, and the device has the advantages of simplicity and convenience.
In one embodiment, HPV lysis is achieved using a centrifugal microfluidic chip. Specifically, a cylindrical small magnet with a diameter of 4mm and a height of 2mm is preset in a stirring chamber of the centrifugal microfluidic chip to serve as a magnetic absorption member, and small glass beads for realizing a grinding effect are preset in the stirring chamber to serve as grinding bodies, wherein the diameter of the small glass beads is 100-600 μm. A permanent magnet is fixed below the centrifugal microfluidic chip, the center of the permanent magnet is closer to the center of the circle, and the stirring cavity is at least partially or totally deviated from the center of the circle. And adding cervical brush eluent into the stirring chamber to obtain a sample for HPV detection, rotating the centrifugal microfluidic chip at the speed of 300rpm, and driving the small glass beads to grind the cervical brush eluent in the stirring chamber for 10 minutes by the back and forth movement of the small magnet at the moment, so that the mixing and the cracking of cells, tissues and HPV viruses in the sample are realized, and the DNA in the sample is released. When the rotational speed of the centrifugal microfluidic chip is increased to 800rpm, at this time, even if the stirring chamber is directly above the permanent magnet 160, the magnetic force is insufficient to attract the small magnet to the end of the stirring chamber near the center of the circle against the centrifugal force. At this time, the small magnet cannot move back and forth in the stirring chamber, and the cracking effect is avoided. The structure is simple, and the control is simple and convenient; without requiring complex instrumentation.
Further, in one of the embodiments, the chip substrate is PMMA, PDMS, PC, ABS, COC or COP. In one embodiment, the chip substrate has a cylindrical structure or the chip substrate has a cylindrical structure. In one embodiment, the chip substrate is further provided with an enrichment chamber, the enrichment chamber and the stirring chamber are separated by a filter membrane, the enrichment chamber is located above the stirring chamber, the enrichment chamber is used for enriching a sample in the stirring chamber by centrifugation and only remaining supernatant in the enrichment chamber, and the liquid injection hole is communicated with the enrichment chamber and is communicated with the stirring chamber by the enrichment chamber. By means of the design, effective substances in a sample are enriched in the stirring cavity, so that the stirring and mixing effects are improved, and the matching and the cracking effects are improved.
In one embodiment, a nucleic acid analysis system includes the centrifugal microfluidic chip of any of the embodiments. According to the nucleic acid analysis system, the ingenious design that the magnetic adsorption piece is movably arranged in the stirring cavity is adopted, and on one hand, when the magnetic adsorption piece moves to a position far away from the magnetic piece, the magnetic adsorption piece moves to a position far away from the rotation center of the stirring cavity under the action of centrifugal force; on the other hand, when the magnetic adsorption piece moves to a position close to the magnetic piece, the magnetic adsorption piece moves to a position of the stirring cavity close to the rotation center under the action of magnetic adsorption; therefore, the ideal mixing effect is realized, and the cracking effect is stable and effective on the premise of simplifying the structure and needing no complex instruments and equipment.
It should be noted that, other embodiments of the present application further include a centrifugal microfluidic chip and a nucleic acid analysis system that are formed by combining the technical features of the above embodiments.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples only represent a few embodiments of the present application, which are described in more detail and are not to be construed as limiting the scope of the claims. It should be noted that it would be apparent to those skilled in the art that various modifications and improvements could be made without departing from the spirit of the present application, which would be within the scope of the present application. Accordingly, the scope of the present application is to be determined by the following claims.
Claims (10)
1. A centrifugal microfluidic chip, comprising: the chip comprises a chip substrate, a magnetic absorption part and a magnetic part;
the chip substrate is also provided with a stirring chamber and a liquid injection hole communicated with the stirring chamber;
the magnetic absorption part is accommodated in the stirring cavity;
the chip substrate is provided with a rotation center, the magnetic piece is arranged outside the chip substrate and positioned at the position of the stirring cavity close to the rotation center, and the magnetic piece is used for enabling the magnetic absorption piece to move to the position of the stirring cavity close to the rotation center under the action of magnetic absorption when the distance between the magnetic piece and the magnetic absorption piece is smaller than a preset value;
the preset value is set according to the balance relation between the magnetic force of the magnetic field and the centrifugal force of the centrifugal field, and the magnetic adsorption piece moves back and forth by utilizing the balance relation between the magnetic force of the magnetic field and the centrifugal force of the centrifugal field.
2. The centrifugal microfluidic chip according to claim 1, wherein the magnetic member is a magnetic body and the magnetic absorbing member is a ferromagnetic body matching the magnetic body.
3. The centrifugal microfluidic chip according to claim 2, wherein the ferromagnetic body is provided with a protective outer layer.
4. The centrifugal microfluidic chip according to claim 1, wherein the magnetic member is a ferromagnetic body and the magnetic adsorbing member is a magnetic body matching the ferromagnetic body.
5. The centrifugal microfluidic chip according to claim 1, further comprising a grinding body disposed in the stirring chamber.
6. The centrifugal microfluidic chip according to claim 1, further comprising a support member provided on the chip base body and not rotating with the chip base body, wherein the magnetic member is fixedly provided on the support member.
7. The centrifugal microfluidic chip according to claim 1, wherein the magnetic member is provided separately from the chip substrate.
8. The centrifugal microfluidic chip according to any one of claims 1 to 7, wherein the number of the magnetic adsorbing elements and the stirring chambers is plural, and each stirring chamber accommodates one magnetic adsorbing element therein.
9. The centrifugal microfluidic chip according to claim 8, wherein the stirring chambers are identical or differently shaped.
10. A nucleic acid analysis system comprising a centrifugal microfluidic chip according to any one of claims 1 to 9.
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| CN110420604B (en) * | 2019-08-30 | 2024-08-23 | 东莞东阳光医疗智能器件研发有限公司 | Disc chip magnetic particle moving device, detecting device and moving control method thereof |
| CN110938522A (en) * | 2019-12-13 | 2020-03-31 | 大连海事大学 | Portable bacteria cracking device based on centrifugal microfluidic technology and use method thereof |
| CN113617400B (en) * | 2020-05-06 | 2025-03-04 | 苏州国科均豪生物科技有限公司 | Centrifugal microfluidic chip |
| CN111548929A (en) * | 2020-05-29 | 2020-08-18 | 天津诺迈科技有限公司 | A centrifugal microfluidic chip for cell lysis and sample homogenization |
| CN115870026B (en) * | 2023-01-04 | 2025-04-08 | 南京师范大学 | A device and method for large-scale production of highly monodisperse droplets |
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| CN106268476A (en) * | 2016-08-05 | 2017-01-04 | 安庆庆荣企业管理咨询有限责任公司 | Magnetic stir bar |
| CN108970655A (en) * | 2018-06-04 | 2018-12-11 | 东莞东阳光科研发有限公司 | A kind of magnetic bead transfer device of micro-fluidic chip |
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| CN106268476A (en) * | 2016-08-05 | 2017-01-04 | 安庆庆荣企业管理咨询有限责任公司 | Magnetic stir bar |
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